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Consequent artefacts, such as blurring and elongation of features in the missing wedge direction, deteriorate the quality of the reconstruction and make quantification difficult (e.g. [12]).
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However, we are aware that these tomograms are still affected by the missing wedge along the z direction, as pointed out by the reviewers.
DOI: http://dx.doi.org/10.7554/eLife.03600.008 The orientation of our crystals prevented the full sampling of reciprocal space leading to systematic incompleteness (missing wedge or missing cone).
Although such tomograms gave us clear hints about the overall arrangement of septin cables and revealed clear differences between septin-Gic1 and septin-Gic1-Cdc42-GppNHp, the missing wedge artifacts caused the broadening of the septin filaments, making their exact tracing difficult in all slice-directions.
Despite the recent progress in the field of reconstruction and regularization techniques, the missing wedge artefacts remain untreated in most current reconstruction workflows in structural biology.
Fig. 5 Missing wedge reconstruction, processed with NAD. a A WBP reconstruction of aligned, noiseless projections with a 28 ° missing wedge.
Fig. 10 Missing wedge reconstruction processed with Weka. a The perfectly aligned, noiseless 28 ° missing wedge reconstruction from Fig. 5. b The same reconstruction but processed with Trainable Weka Segmentation.
All tomograms are affected by missing wedge artifacts.
to a popular constrained correlation score with missing wedge correction (Förster et al., 2008).
The missing wedge effect leads to distortions of the density maps in real spaces.
We did not claim that there are no missing wedge artifacts.
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